KR20090016707A - How to treat autoimmune diseases using TACIBIV fusion molecules - Google Patents

Abstract

In various embodiments, the present invention provides methods and compositions for treating autoimmune diseases including rheumatoid arthritis, wherein the methods of the present invention comprise administering a TACI-Ig fusion molecule to a patient in need thereof, for example. It includes. In one embodiment, the TACI-Ig fusion molecule is administered in an amount sufficient to delay, inhibit or inhibit the proliferation-inducing function of BIyS and APRIL.

Description

METHODS FOR TREATING AUTOIMMUNE DISEASES USING A TACI-IG FUSION MOLECULE}

Cross Reference to Related Application

This application claims priority to US Provisional Application No. 60 / 747,270, filed May 15, 2006, the contents of which are incorporated herein by reference.

Field of invention

In various embodiments, the present invention relates to methods and compositions for treating autoimmune diseases or disorders of the immune system comprising administering a TACI-Ig fusion protein that blocks the function of growth factors of the TNF family. .

Background of the Invention

BlyS Ligand / Receptor Family

Three receptors with unique binding affinity for two growth factors, BlyS (B-lymphocyte stimulator) and APRIL (proliferation-inducing ligand), ie TACI (transmembrane activator or calcium-regulated cyclo Filin ligand-interacters), BCMA (B-cell maturation antigen) and BAFF-R (receptor for B-cell activating factor, belonging to the TNF family) have been identified (Marsters et al. Curr Biol 2000; 10 (13) : 785-788; Thompson et al. Science 2001; 293: 21 08-2111). TACI and BCMA bind to both BLyS and APRIL, but BAFF-R is believed to only bind BLyS with high affinity (Marsters et al. Curr Biol 2000; 10 (13): 785-788; Thompson et al. Science 2001; 293: 21 08-2111). As a result, it is believed that BLyS can transmit signals through all three receptors, while APRIL can only transmit signals through TACI and BCMA. In addition, a cyclic heterotrimeric complex of BLyS and APRIL (3 proteins containing one or two copies of BLyS and APRIL, respectively) is collected from a patient with systemic immune-based rheumatic disease In serum samples and was found to induce B-cell proliferation in vitro (Roschke et al. J Immunol 2002; 169: 4314-4321). Of the Ig-fusion proteins for all three receptors, only TACI-Fc5 could block the biological activity of the heterotrimeric complex (Roschke et al. J Immunol 2002; 169: 4314-4321).

BLyS and APRIL are potent stimulators of B-cell maturation, proliferation and survival (Gross et al. Nature 2000; 404: 995-999. Gross et al. Immunity 2001; 15 (2): 289-302. Groom et al J Clin Invest 2002; 109 (1): 59-68). BLyS and APRIL may be required for the persistence of autoimmune diseases, especially autoimmune diseases involving B-cells. Transgenic mice engineered to express high levels of BLyS exhibit immune cell disorders and symptoms similar to those observed in patients with systemic lupus erythematosus (Cheson et al. Revised guidelines for diagnosis and treatment. Blood 1996; 87: 4990-4997. Cheema et al. Arthritis Rheum 2001; 44 (6): 1313-1319). Similarly, increased levels of BLyS / APRIL were measured in serum samples taken from SLE patients and other patients with various autoimmune diseases such as rheumatoid arthritis (Roschke et al. J Immunol 2002; 169: 4314-4321). ; Mariette X., Ann Rheum Dis 2003; 62 (2): 168-171; Hahne et al. J Exp Med 1998; 188 (6): 1185-1190), which are BLyS and / or APRIL and B-cell mediated diseases Extends the association of humans from animal models to humans.

Rheumatoid arthritis

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by nonspecific and generally symmetrical inflammation of the surrounding joints and potentially causes gradual destruction of joints and periarticular structures. About 1% of the population suffers from this disease, and women are two to three times more frequent than men. The exact etiology of the disease is not known, but strong evidence suggests that RA is an autoimmune disease. Several auto-antibodies are associated with RA, examples of which are antibodies to rheumatoid factor, anti-nuclear factor and native collagen type II and citrulled peptides, which are often associated with the more severe disease.

Prominent immunological abnormalities that may be important in the development of RA include immune complexes found in articular fluid cells and vasculitis. Contributing to this complex are antibodies (eg, RF) produced by plasma cells and T helper cells capable of penetrating synovial tissue and producing pro-inflammatory cytokines. Macrophages and their cytokines (eg TNF, GMCS-F) are also abundant in the diseased synovial membrane. Increased levels of adhesion molecules contribute to the retention and retention of inflammatory cells in synovial tissue. Increased macrophage derived lining cells are also prominent with some lymphocytes.

The role of T cells in the onset of RA is well established, while the role of B cells is less well known. Nevertheless, B cells play a number of potential roles in the development of RA, which act as antigen-providing cells, secrete proinflammatory cytokines, produce rheumatoid factor auto-antibodies and activate T cells. It involves doing. The potential role of B cells is further supported by the positive results of clinical trials testing rituximab, a monoclonal antibody directed against CD20 in RA patients, when given with methotrexate or cyclophosphamide. Most noticeable. These results suggest that B cells are an appropriate target for therapeutic intervention in RA.

Confirmed therapeutic agents for RA include disease modifying anti-rheumatic drugs (DMARDs), for example hydroxychloroquine, sulfasalazine, methotrexate, leflunomide, rituximab, infliximab, aza Thioprine, D-penicillamine, Gold (oral or intramuscular), minocycline and cyclosporin, corticosteroids such as prednisone and non-steroidal anti-inflammatory drugs (NSAIDs). Such therapeutic agents are generally nonspecific and frequently associated with serious side effects and do not significantly affect the progression of joint destruction. As a result, the development of new methods for treating RA is a necessary requirement in the art.

Summary of the Invention

In various embodiments, the present invention relates to a method for treating an autoimmune disease. Illustratively, the methods of the present invention comprise administering to a patient a composition comprising a human immunoglobulin-constant domain and a TACI extracellular domain or a fragment thereof that binds BlyS and / or APRIL.

In another embodiment, the present invention treats an autoimmune disease comprising RA using a TACI-Ig fusion molecule comprising a TACI extracellular domain or any fragment thereof that retains the ability to bind BlyS and / or APRIL. It includes how to do it.

In another embodiment, the invention administers to a patient in need of a RA treatment a fusion molecule comprising an effective amount of a human immunoglobulin-constant chain and a fragment of the TACI extracellular domain or TACI extracellular domain that binds BlyS and / or APRIL. Methods of treating RA, including In one embodiment, the fragment of the extracellular domain of TACI comprises one or two cysteine repeat motifs. In another embodiment, the fragment is a fragment comprising amino acids 30-110 of the extracellular domain of TACI. In another embodiment, the fragment is a fragment comprising amino acids 1-154 (SEQ ID NO: 1) of the extracellular domain of TACI.

In another embodiment, the invention is a fusion polypeptide comprising a human immunoglobulin- constant domain Fc5 having the sequence set forth in SEQ ID NO: 2 and a TACI extracellular domain having the sequence set forth in SEQ ID NO: 1 A method of treating RA by administering a composition comprising TACI-Fc5 to a patient.

In another embodiment, the invention binds to a human immunoglobulin-constant domain having a sequence set forth in SEQ ID NO: 2 and BlyS and / or APRIL and at least about 50%, at least about 60% with SEQ ID NO: 1 At least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 99%. A method of treating RA by administering the composition to a patient.

Other autoimmune diseases can be treated by the methods of the invention by administering to the patient a fusion polypeptide comprising a human immunoglobulin-constant chain and a fragment of the TACI extracellular domain or TACI extracellular domain that binds BlyS and / or APRIL. Can be. Such autoimmune diseases include, but are not limited to, systemic lupus erythematosus (SLE), Graves' disease, type I and II diabetes mellitus, multiple sclerosis, Sjogren's syndrome, scleroderma, glomerulonephritis, transplant rejection, for example organ and tissue allografts. And xenograft rejection and graft versus host disease.

In one embodiment, the methods of the present invention comprise administering to the RA patient an TACI-Ig fusion molecule in an amount from about 0.01 mg to about 10 mg per kg body weight of the patient. The TACI-Ig fusion molecule may be repeatedly administered at predetermined intervals. By way of example, molecules may be administered seven or more times over a 12 week interval. Following this initial treatment with a TACI-Ig fusion polypeptide, the polypeptide may be administered once every other week (every other week) for at least two additional weeks, for example, the polypeptide may be administered at two weeks for an additional two to thirty weeks. It may be administered once. In addition, the polypeptide may be administered weekly or daily.

According to the methods of the invention, the TACI-Ig fusion polypeptide may be administered to the RA patient by subcutaneous, oral or intravenous routes, and with other agents. Such agents include, but are not limited to, DMARDs such as hydroxychloroquine, sulfasalazine, methotrexate, leflunomide, rituximab, infliximab, azathioprine, D-penicillamine, gold (oral or intramuscular). ), Minocycline and cyclosporine, corticosteroids such as prednisone, NSAIDs, cytokines, anti-cytokines and interferons.

1 shows a dose-increasing decision tree for TACI-Fc5 treatment.

2 illustrates a patient body region that can be used for subcutaneous injection of TACI-Ig molecules.

Detailed description of the invention

In various embodiments, the present invention relates to a method of treating an autoimmune disease in a patient by inhibiting the interaction of BlyS and / or APRIL with their receptors. The patient can be a mammal, eg a human. In one embodiment, the methods of the invention comprise 1) a polypeptide comprising a domain at least partially identical to a TACI extracellular domain or a fragment thereof that binds BlyS and / or APRIL and 2) an inhibitor comprising a human immunoglobulin constant chain Use In one embodiment, the methods of the present invention comprise at least about 50%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80% with human immunoglobulin constant chains and TACI extracellular domains. At least about 85%, at least about 90%, at least about 95% or at least about 99% of the fusion molecules are used that include any polypeptide having sequence identity. U.S. Pat.Nos. 5,969,102, 6,316,222 and 6,500,428 and U.S. Patent Applications 09 / 569,245 and 09 / 627,206, the teachings of which are incorporated herein by reference in their entirety, include TACI ligands, including BlyS and APRIL, as well as extracellular domains of TACI. The sequences for specific fragments of TACI extracellular domains that interact are described. One exemplary fragment of the extracellular domain of TACI includes one or two cysteine repeat motifs. Another exemplary fragment is a fragment comprising amino acids 30-110 of the extracellular domain of TACI or a fragment thereof. Another exemplary fragment is a fragment or fragment thereof comprising amino acids 1-154 (SEQ ID NO: 1) of the extracellular domain of TACI.

Other fusion molecules useful in the methods of the present invention include fusion polypeptides between human immunoglobulin constant chains and complete TACI extracellular domains or their orthologs or extracellulars capable of binding human immunoglobulin constant chains with BlyS and APRIL ligands. There is a fusion polypeptide between any fragment of the TACI domain. Any one of the fusion molecules used in the methods of the present invention may be referred to as a TACI-Ig fusion molecule.

TACI-Fc5 is one of the TACI-Ig fusion molecules useful in the methods of the present invention. TACI-Fc5 is a recombinant comprising an extracellular ligand-binding portion (SEQ ID NO: 1) of receptor TACI from amino acid about 1 to about 154 amino acids and Fc5 (SEQ ID NO: 2), which is a modified Fc portion of human IgG Fusion polypeptides. Other TACI-Ig molecules useful in the methods of the present invention may bind to polypeptides having SEQ ID NO: 2 and BlyS, at least about 50%, at least about 60%, at least about 65%, There are fusion molecules comprising at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95% or at least about 99% identical polypeptides.

Embodiments of the invention include methods of using TACI-Ig fusion molecules to treat RA. Other autoimmune diseases that can be treated by the methods of the invention include systemic lupus erythematosus (SLE), Graves' disease, type I and II diabetes, multiple sclerosis, Sjogren's syndrome, scleroderma, glomerulonephritis, transplant rejection, eg For example, organ and tissue allograft and xenograft rejection, graft-versus-host disease or any of those that can be treated by reducing the number of circulating mature B-cells, immunoglobulin-secreting cells and soluble immunoglobulins associated with these diseases. There is an autoimmune disease outside.

Embodiments also include methods of treating by administering to a patient a fusion molecule comprising a polypeptide comprising any fragment of a TACI extracellular domain capable of binding a human immunoglobulin-constant domain and BlyS and / or APRIL.

The TACI-Ig fusion molecule can be administered to a patient according to any suitable route of administration, including but not limited to the oral, intravenous or subcutaneous route.

TACI-Ig formulations useful in the methods of the invention can be prepared and stored as freeze, sterile, isotonic solutions. Such formulations may include other active ingredients and excipients such as sodium chloride, phosphate buffer and sodium hydroxide or O-phosphate, pH 6.0. TACI-Ig formulations may be administered to the patient in combination with other agents. Such agents include, but are not limited to, DMARDs such as hydroxychloroquine, sulfasalazine, methotrexate, leflunomide, rituximab, infliximab, azathioprine, D-penicillamine, Gold ) (Oral or intramuscular), minocycline and cyclosporin, corticosteroids such as prednisone, NSAIDs, and pain medications. The method of the present invention can be used in conjunction with other methods of treating autoimmune diseases. Such other treatments include, but are not limited to surgery, acupuncture, physiotherapy and gene therapy. TACI-Ig formulations may be administered before, concurrently with or after other methods of treatment.

TACI-Fc5 has been shown to inhibit BlyS activation of B cell proliferation in vitro. Treatment of mice with TACI-Fc5 has minimal effect on B cell precursors in bone marrow and other cell lineages including peripheral blood T cells, monocytes and neutrophils, and partially blocks B cell development. Transgenic mice engineered to overexpress soluble forms of TACI receptors in the blood produce fewer mature B cells and exhibit reduced levels of circulating antibodies. TACI-Fc5 transgenic mice had normal numbers of cells in the thymus, bone marrow and mesenteric lymph nodes. There was no significant difference in the T cell population in the thymus, lymph nodes and spleen (Gross et al. Immunity 2001; 15 (2): 289-302).

In addition, TACI-Ig can inhibit antigen-specific antibody production in the immune response of mice, whether administered during a primary or secondary response to an antigen. In this study, no effect on T cell responses to ex vivo antigen challenge was observed. In animal models of systemic lupus erythematosus, treatment with TACI-Ig fusion proteins has been effective in limiting the onset and progression of the disease (Gross et al. Nature 2000; 404: 995-999). Similarly, in mouse models of collagen-induced arthritis, TACI-Ig could inhibit the development of collagen-specific antibodies and reduce both the incidence of inflammation and the incidence of disease (Gross et al. Immunity 2001; 15 (2): 289-302).

Compositions comprising a TACI-Ig fusion molecule may be administered to a patient once or repeatedly over a period of time. For example, a patient may receive a single subcutaneous injection of TACI-Ig molecules, after which the patient's disease may be monitored. Patients who exhibit amelioration or at least stabilization of the disease may receive repeated administration of the TACI-Ig fusion molecule for an additional period of time. The additional period of time may be from about 2 weeks to about 30 weeks. For example, a patient may receive three administrations of the TACI-Ig fusion molecule for a four week period. In addition, the patient may receive seven administrations of the TACI-Ig fusion molecule for a 12 week period. Administration of the TACI-Ig molecule may be in units of daily, every other day, weekly, biweekly, monthly, bimonthly, and the like.

The TACI-Ig fusion molecule is administered to a patient in an amount effective to treat the patient's disease. In one embodiment, the term “treatment” in connection with a given disease or disorder includes, but is not limited to, inhibiting the disease or disorder, eg, arresting the occurrence of the disease or disorder; Alleviating the disease or disorder, for example causing regression of the disease or disorder; Or alleviating a disease caused or caused by a disease or disorder, for example, alleviating, preventing, or treating the symptoms of a disease or disorder. In another embodiment, the amount can range from 0.01 mg to 10 mg per kg of body weight of the patient. The optimal amount for treatment with a TACI-Ig fusion molecule can be found by using the diagram in FIG. 1, which is described in more detail in Example 5.

Fusion TACI-Ig molecules can be delivered in any suitable manner. In one embodiment, the molecule is delivered by intraperitoneal injection. In another embodiment, the intraperitoneal injection is via subcutaneous injection. In another embodiment, the intraperitoneal injection is administered into the anterior abdominal wall. If more than one injection is required to administer the dosage, the injections may be administered at several centimeter intervals and relatively close in time, for example as reasonably as close as possible. For repeated drug administration, the site of administration on the anterior abdominal wall can be circulated or alternating. Typical sites for subcutaneous injection into the anterior abdominal wall are shown in FIG. 2, which include the right upper outer region, lower left outer region, lower right outer region, upper left outer region, median lower region as well as the left and right thighs and There is a brachial arm (FIG. 2). In addition, the TACI-Ig fusion molecules of the invention may be delivered via intravenous injection or oral route in the form of tablets, caplets, liquid compositions or gels and the like.

Example 1

TACI-Fc5 Pharmacology, Toxicology and Pharmacodynamic Tests in Experimental Animal Models.

TACI-Fc5 was evaluated in a host resistance model, which provides an opportunity to directly assess the functional reserve of the immune system. Mice were challenged with influenza virus during TACI-Fc5 treatment. Dexamethasone, used as a positive control, resulted in enhanced and long-term viral infection. Single administration of TACI-Fc5 by the subcutaneous (SC) pathway reduced circulating B cells, total IgG and IgM, and influenza-specific IgG and IgM, but did not reduce the animal's ability to eliminate viral infection.

Important safety pharmacology studies have shown that TACI-Fc5 does not induce significant changes in the nervous, respiratory and cardiovascular systems of mice or monkeys at SC doses below 80 mg / kg. In mice only, slightly increased hyper-alertness and motor activity were observed at 80 mg / kg, suggesting a mild and transient stimulating effect, and no-observed effect level (NOEL). ) Was 20 mg / kg.

When administered to mice as a single dose by the intravenous (IV) or SC route, TACI-Fc5 is fatally detectable or systemic or topical in animals at or below the highest technically acceptable dose of 1200 mg / kg. Did not induce abnormal effects.

Administration of TACI-Fc5 as a single dose by the SC route at a dose level of 240 mg / kg did not result in lethal or any significant toxic effects.

Based on the results obtained two or four weeks after the administration of TACI-Fc5 by the subcutaneous route to mice at doses of 5, 20 and 80 mg / kg every 2 days, the compound was 80 mg / kg It can be concluded that tolerability in these species is good at the following dosages. Treatment-related changes specific to the immune system were seen at all doses. These changes included reductions in total and mature B cell numbers and IgG and IgM serum levels. Immunohistochemical tests performed on the spleen and lymph nodes confirmed depletion specific to B cells, but the T cell number did not change. In some cases all these time- and dose-related changes were considered as exaggerated pharmacological effects as expected in reactive species after very high doses of TACI-Fc5. Taken together, these effects were observed two and four weeks after treatment, with no significant signs of progressing over time. This effect appeared almost completely reversible except for the reduced B cell count 4 weeks after treatment was stopped.

To confirm B-cell regulatory reversibility, additional experiments in mice were performed at doses of 5 and 20 mg / kg administered every 2 days for 4 weeks, providing a longer recovery period. Recovery of total and mature circulating B cells occurred 2 months after stopping for 5 mg / kg and 4 months after 20 mg / kg. Moreover, the injection induced a slight increase in inflammatory change at all injections compared to the vehicle control at the injection site.

Subcutaneous administration of TACI-Fc5 in monkeys did not induce significant signs of toxicity at any of the test doses 5, 20 or 80 mg / kg administered every three days when administered for four consecutive weeks.

Local tolerability was deemed satisfactory at doses up to and including the highest dose tested. Dose related and reversible slight or moderate changes (mainly perivascular monocytes and eosinophil infiltration) of inflammatory origin were induced, but were primarily considered to be related to the local presence of exogenous proteins. At high doses only, several animals showed slight or moderate subacute inflammation associated with cyst formation in one of them.

Lymphocyte subset measurements showed a decrease in circulating B-cell numbers as well as histological depletion of the splenic follicle margin (known as the B-cell dependent region) and a decrease in total IgG and IgM serum levels. This was considered as a result of the pharmacokinetic properties of TACI-FcS as revealed by in vitro and in vivo pharmacology experiments. The extent was excessive, as expected in toxicology studies where animals were intentionally administered high doses of the test compound. Although low serum IgG and IgM levels and splenic lymphocyte depletion showed a clear trend for recovery within a one-month pause period allowed, total and mature circulating B cells did not show similar behavior, which required longer time to recover. Indicates.

At the end of the treatment period (week 4), males and females of the high dose group (80 mg / kg) showed a slight but statistically significant decrease in mean total protein values compared to the control. A slight trend of decline was also observed at the same dose at week 2 and at the end of the recovery period.

Serum protein changes in high dose females at the end of the dosing period included a decrease in globulin and an increase in albumin ratio and alpha-1-globulin fraction. The alpha-1-globulin fraction was also found to be higher than the control in group 3 females (20 mg / kg).

Immunogenicity of TACI-FcS was found in both mice (only a few females showed low levels of circulating binding antibodies during and after treatment) and monkeys (low levels were found after recovery in several animals). Was low, and there was no evidence of neutralizing antibodies in each species.

Embryo-fetal development by testing TACI-Fc5 in a series of standard in vivo trials (embryo-fetal development studies were conducted on the sc route at doses of 5, 20 and 80 mg / kg every 2 days during organogenesis. Performed in female mice and rabbits) and reproductive and reproductive toxicology (reproduction testing was carried out by the sc route at doses of 5, 20 and 80 mg / kg before and during mating and every 2 days until implantation period). (Done in male and female mice).

Reproduction tests in mice showed a dose related increase in preimplantation and postimplantation loss after exposure to 20 and 80 mg / kg of TACI-Fc5 every 2 days compared to the control group.

Evaluation of the data obtained in mice of the embryo-fetal development study indicated that no embryotoxic effect was observed at any dose and no compound related fetal malformation was induced.

For rabbits, embryo-fetal development studies showed that treatment induced dose-related lower weight gain and lower food consumption in pregnant animals treated at 20 or 80 mg / kg every 2 days. The maternal change was associated with increased resorption rate and lower fetal body weight at two higher doses.

These results suggest a possible effect of TACI-Fc5 on implantation of mouse blastocysts in the uterus. The observed effects of TACI-Fc5 on maternal weight gain and food consumption are believed to be responsible for the observed effects on litter viability in rabbits exposed to 20 or 80 mg / kg every 2 days during organogenesis. There is no direct toxicity of TACI-Fc5. There were no malformations due to TAI-Fc5 treatment in these two animal species.

In addition, histological examinations of male and female gonads and adjuvant reproductive organs were carried out as a two-week and one-month toxicity study by the sc route in mice and monkeys administered every 2 or 3 days, respectively, without evidence of the effects associated with TACI-Fc5 treatment. Was performed.

Topical tolerability studies in rabbits showed that localized tolerability was good when the TACI-Fc5 formulation was injected by the subcutaneous route into rabbits at a dose of 70 mg / mL.

Single dose pharmacokinetic studies in male mice by the IV and SC routes were performed by intravenous route at doses of 1 mg / kg in mice or by subcutaneous route at doses of 1, 5 and 15 mg / kg. It was.

The time to maximum absorption (t _max ) was calculated between 4 and 16 hours and t _1/2 was calculated to be about 40-50 hours.

An infusion-like profile was observed during the first 30 minutes after IV bolus administration, after which TACI-Fc5 was removed from the body with a removal half-life of 44 hours. After subcutaneous administration, the ratio between the AUC (area curve area) obtained at three doses of 1, 5 and 15 mg / kg was 1: 5: 8 for a dose ratio of 1: 5: 15, which is a high dose. Implies a loss of dose proportionality in.

 The bioavailability of TACI-Fc5 by the subcutaneous route was 76 and 89% at doses of 1 and 5 mg / kg, but lower than expected at 15 mg / kg in mice (0,42; intravenous 1 mg / kg). Volume is calculated for the volume). Since the apparent elimination half-life did not change, the lower bioavailability observed at higher doses could be explained by an increase in both elimination and distribution volume, or more certainly reduced absorption due to the formation of deposits at the injection site. It can be explained by.

Single dose pharmacodynamic studies in male monkeys by the IV and SC routes were administered by the intravenous route at a dose of 1 mg / kg or by the subcutaneous route at doses of 1, 5 and 15 mg / kg. It was performed in cynomolgus monkeys.

Six male monkeys were divided into two groups of three and received two doses, each administered by a two week wash-out period. Period 1 treatments were 1 mg / kg IV (group 1) and 1 mg / kg SC (group 2), and period 2 treatments were 5 mg / kg SC (group 1) and 15 mg / kg SC (group 2).

The time to maximum absorption (t _max ) was calculated between 6 and 8 hours and t _1/2 was calculated to be about 120 to 190 hours.

An infusion-like profile was observed in two of the three monkeys during the first 15 minutes after IV bolus administration, after which TACI-Fc5 was removed from the body with an elimination half-life of 179 ± 29 hours. The distribution volume Vss at steady state was 382 ± 82 mL / kg, which is close to the intracellular fluid volume.

 After subcutaneous administration, AUC for dose proportionality was good, ie 216, 1182 and 2732 h μg / mL for SC doses of 1, 5 and 15 mg / kg. TACI-Fc5 bioavailability (calculated for 1 mg / kg IV dose) by the subcutaneous route was 0.92, 1.02 and 0.77 at low, medium and high doses. Thus, TACI-Fc5 was absorbed almost completely by the subcutaneous route.

Low levels of TACI-Fc5 were found in pre-dose samples for period 2 for all six monkeys (dose 1 mg / kg by period IV or SC route and 5 or 15 mg / kg of period 2, respectively). Between administrations) because only two half-lives have elapsed during the two-week washout period, and the number of such half-lives is insufficient for complete removal of the administered compound (requires five half-lives). However, the AUC contribution of the previous dose could be estimated to represent only about 2% of the total AUC in period 2.

IgG serum levels showed a 10.2% decrease after IV administration. The 15 mg / kg SC dose showed a slightly higher effect, but no difference was observed between the 1 and 5 mg / kg SC doses (8.6%, 8.4% and 12.3% after 1, 5 and 15 mg / kg, respectively). decrease). IgM serum levels showed a 18.0% decrease after IV administration. No differences were observed between the three SC doses (23.5%, 23.0% and 24.2% reductions after 1, 5 and 15 mg / kg administration, respectively).

Example 2

Tolerance of TACI-Fc5 in Healthy Volunteers

The first Phase I study of TACI-Fc5 is currently being completed. This is a double-blind, placebo-controlled, incremental dose study to investigate the safety, pharmacodynamics, and pharmacokinetics of a single dose of TACI-Fc5 administered subcutaneously to healthy male volunteers. A summary of the study design is provided below along with a summary of the available data.

To investigate the safety, pharmacodynamics, and pharmacokinetics of a single dose of TACI-Fc5 administered subcutaneously to healthy male volunteers, TACI-Fc5 was administered to healthy volunteers in a double-blind, placebo-controlled, escalating sequential dosing study.

Four groups of subjects were recruited. In each dose group, one subject was randomly selected to receive a placebo injection and all the other subjects received TACI-Fc5. After leaving the study site 24 hours after dosing, the subjects were outpatient for a 7 week scheduled assessment. Systemic and local tolerability of TACI-Fc5 physical examination results, injection site pain, local tolerability reactions (redness, swelling, bruising and itching), vital signs, 12-lead (lead) Monitored by ECG (electrocardiogram), safety laboratory evaluation and recording of adverse events.

Pharmacokinetic and pharmacokinetic markers were monitored throughout the 7 week post administration period. The pharmacokinetic effects of TACI-Fc5 were monitored using a number of markers, which include lymphocyte subsets (plasma cells (CD138 +), immature B cells (CD19 +, IgD-), mature B cells (CD19 +, IgD +), T-helper cells (CD5 +, CD4 +), cytotoxic T-cells (CD5 +, CD8 +), total T-cells (CD5 +)), free BlyS, BLyS / TACI-Fc5 complex, IgG, IgM, anti- TACI-Fc5 antibody.

Dose escalation was induced by an algorithm within the study protocol based on a review of the data at 3 weeks post dose. In four groups, group 1 received 2.1 mg, group 2 received 70 mg, group 3 received 210 mg, and group 4 received 630 mg.

Results: Healthy male volunteers received a single subcutaneous dose of TACI-Fc5 at doses ranging from 0.03 mg / kg to 9 mg / kg. Safety and tolerability data were used in conjunction with FACS analysis of lymphocyte subsets at week 3 to induce dose escalation between cohorts. Four cohorts were studied as shown in Table 1.

Table 1: Repartitioning of Volunteers

^a nominal weight of 70 kg was assumed at each dose level and the subject received a standardized dose.

^b Due to the dilution error, the dose administered to cohort 1 was 10 times lower than the planned dose (0.3 mg / kg). This error was confirmed upon review of pharmacodynamic data from Cohort 1, but this confirmation was made after administration to Cohort 2.

^c Include one subject for placebo.

1 volunteer gave up before ^d injection.

Demographic baseline characteristics are summarized for cohorts by cohort and in total, and are shown in Table 2.

Table 2: Demographic Characteristics

Taken together, mean +: SD age was 30.7 + 7.4 years and mean BMI was 24.8 kg / m2. All volunteers were white males.

TACI-Fc5 was well tolerated in all groups. Physical examination revealed no obvious effects on vital signs or 12-lead ECG.

Table 3: List of adverse events resulting from treatments reported to date

Transient redness and swelling were observed at the site of administration of some subjects, and redness affected all subjects in cohorts 3 and 4. Although the incidence of injection site reactions is believed to increase in higher dose groups, it is believed to be associated with increased injection volume (and frequency).

Adverse events were reported following 48 treatments within 7 weeks of dosing. Most of them (44 cases, 91.7%) were mild, and the others (4 cases, 8.3%) were moderate. There were no serious adverse events or serious adverse events during this period. There was no obvious relationship between the dose of TACI-Fc5 administered and the incidence, intensity or established relationship of adverse events. Adverse events reported to date are summarized in Table 3.

It is believed that TACI-Fc5 showed good tolerability at doses up to 630 mg, and no significant safety issues were raised. These data support the intended dose of the proposed subject study.

Non-compartmental analysis of TACI serum concentrations was performed. This preliminary analysis was performed using nominal sampling time. Subjects 2, 6, and 13 had measurable concentrations prior to dosing, so baseline concentrations were subtracted from all post-dosing measurements prior to analysis. Pharmacodynamic parameters after a single subcutaneous administration of 2.1, 70, 210 and 630 mg are summarized in Table 4. Drug concentrations approached the limit of quantification of the assay after 2.1 mg administration of TACI-Fc5, which limits the value of the data at this dose level. At doses of 70 mg and above, T _max (time to maximum absorption) ranged from 16 to 36 hours and total median t _1/2 (calculated from the distal end of the curve) was 303 hours. . In addition, AUC (extrapolated to infinity) and C _max increased higher than in dose proportional mode.

Pharmacokinetic analysis showed a decrease in baseline IgM levels for 7 weeks after one dose of 70, 210 or 630 mg. Although no apparent dose response relationship could be established with a small sample size, the extent of IgM reduction was greatest in the highest dose group. Subjects in the 70 mg dose group were considered to indicate that IgM levels returned to baseline by 7 weeks post-dose. Levels in the higher dose group remained suppressed at this point. There was no apparent effect on the lymphocyte subpopulation measured by IgG levels, or FACS.

During the sampling period the level of BLyS / TACI-Fc5 complex increased proportionally and reached plateau up to about 600 hours post dose. CONCLUSIONS: Human data obtained from healthy male volunteers showed that TACI-Fc5 was well tolerated and safe by subjects at doses up to 630 mg. The nature, incidence and severity of adverse events were comparable between the TACI-Fc5 treatment group and placebo. Physical examination revealed no clinically significant changes in vital signs, 12-lead ECG or safety laboratory parameters. Local tolerability at the site of administration was good. These data support the proposed dose in subjects with BCM.

After a single dose in healthy male subjects, TACI-Fc5 reached T _max between 16 and 20 hours and AUC increased in a dose proportional fashion, but an increase in C _max was higher than in the dose proportional fashion. The median half-life of TACI-Fc5 was about 300 hours. Pharmacokinetic effects were noted for IgM levels at doses of 70, 210 and 630 mg. There was no apparent effect of treatment on lymphocyte subpopulations or IgG after single administration of TACI-Fc5. There are no known or expected risks of specific severity or severity not yet considered in the proposed study protocol.

Example 3

Treatment of RA Patients with TACI-Fc5 Compositions

RA patients are clinically evaluated prior to administration of TACI-Fc5. Baseline assessment is defined as the assessment performed immediately prior to initial administration of TACI-Fc5. The first day of TACI-Fc5 administration is indicated as "day 1". Complete the following procedures before administering the patient the initial dose of TACI-Fc5 dosing: ECG; Vital signs (VS) measurements; Physical examination; Assessment of disease activity (see Example 6); Measurement of height and weight (only for the first dose and subsequent visits); Routine laboratory tests (see Table 5); Calculation of the urine hydroxypyridinolin / lysylpyridinoline (HP / LP) ratio; Free and Total TACI-Fc5 Serum Concentration by Flow Cytometry, Free APRIL and BlyS, Serum Levels of BLyS / TACI-Fc5 Complex, Pharmacodynamic (PK) and Pharmacokinetic (PD) Determination of Cell Number and Anti-TACI Antibody Measurement of (only for the first dose); And lymphocyte subsets (blood split and phenotyped into three tubes by FACS analysis according to Table 6), IgA, IgM, IgG (including sub-typing), anti-citrulized peptide self- Antibodies, rheumatoid factor (IgA-RF, IgM-RF, IgG-RF), C-reactive protein (CRP), TNF-α, IFN-γ, IL-6, IL-1, IL-12, and IL-8 Measurement of blood biomarkers comprising a. In a subset of patients, synovial fluid sampling by arthroscopically induced synovial biopsy or needle puncture is performed, and PK, PD and biomarker assessments are performed in synovial fluid. In a repeated dose cohort, many or all of these measurements are performed immediately prior to subsequent administration of TACI-Fc5.

Table 5: Typical Laboratory Parameters

Blood chemistry hematology γ glutamyl transferase Hematocrit Alanine Traxxamise hemoglobin albumin Average cell hemoglobin Alkaline phosphatase MCHC Aspartate Transaminase Average cell volume Bilirubin-Direct (only if the total bilirubin is outside the normal range) Platelet count Bilirubin-All Red blood cell count calcium Leukocyte count ² Creatinine ESR (erythrocyte sedimentation rate) Glucose (fasting) Urine analysis Weapon Inn pH potassium leukocyte Protein-All Nitrite Electrophoresis of Proteins Glucose Complement 3 Ketone salt Protein ³ Triglycerides ¹ Blood ³ cholesterol coagulation Element Activated partial thromboplastin time Uric acid Prothrombin Time 1: Patients should fast for about 10 hours before measuring triglycerides. 2: White blood cell count should include differential white blood cell count. 3: If protein, blood or white blood cells (RBC or WBC) are present, microscopic examination of the specimen will be performed.

Table 6: Lymphocyte Subsets

tube Antigen markers Tube 1.  CD45 CD3 CD4 CD8 Tube 2.  CD19 msIgG msIgG msIgG (Control) Tube 3.  CD19 CD38 CD27 IgD Tube 1 % and Abs CD45 + / CD3 + (total T-cells)% and Abs CD45 + / CD3 + / CD4 + / CD8- (T-helper cells)% and Abs CD45 + / CD3 + / CD4- / CD8 + (T-cytotoxic / suppressor Cells) Tube 3: % Abs ( CD19 + ) and Abs CD19 + (pan B-cells)% and Abs CD19 + / IgD- / CD27- (immature B-cells) ^* % backcounted using total B cell numbers And Abs CD19 + / IgD + / CD27- (mature B-cells) ^* % and Abs CD19 + / CD27 + / CD38- (memory B cells) ^* % and Abs CD19 + / CD27 + / CD38 + (Ig secretory cells) ( ^* Naive B cells)

At about 6 hours post-dose, the following evaluations are performed except for cohort 6: ECG; VS; And PK and PD sampling (only for single dose cohort).

Treatment with TACI-Fc5 follows the sequential dose-increasing cohort design outlined in FIG. 1. The first cohort is given a 70 mg dose of TACI-Fc5 once a day. Subsequent cohorts are administered up to a maximum dose of 2940 mg based on the dose-increasing algorithm. The second cohort is administered three 70 mg doses of TACI-Fc5 over a 1 month period. The third cohort is given one 210 mg dose of TACI-Fc5. In the fourth cohort, 210 mg doses of TACI-Fc5 were administered three times over a 1 month period. The fifth cohort is given a single 630 mg dose of TACI-Fc5. The sixth cohort received seven doses of 420 mg TACI-Fc5 over a three month period.

Post-dose assessments, including collection of PK, PD and biomarker samples and assessment of safety and disease activity, are performed up to 14 weeks post dose (one dose cohort), up to 18 weeks post dose (repeat dose cohorts 2 and 4) and post dose Complete from all cohorts at predetermined intervals by week 26 (repeat dosing cohort 6). Table 7 lists the post-administration evaluation schedules of cohorts 1, 3 and 5 (single dose cohort) starting on day 2. Table 8 lists the post-dose evaluation schedules of cohorts 2 and 4 starting on the first day after the last dose (day 30). Table 9 lists the post-administration evaluation schedule of cohort 6 starting on the day of dosing of the second dose (Day 15). Injection site reactions, adverse events and accompanying dosing and procedures are measured throughout the study for all cohorts. Adverse event data is obtained based on information provided by the patient at the scheduled or unscheduled study visit after physical examination and by querying the patient.

Adverse events are any annoying medical event in the form of new or worsening signs, symptoms, abnormal laboratory results, or disease, regardless of causality and even when the product under investigation has not been administered (ie, first visit). It is defined as The severity of an adverse event can be assessed using the following definitions: (1) Mild-the patient is aware of the event or symptom but the event or symptom is easily tolerated; (2) moderate-the patient experiences discomfort sufficient to interfere with or reduce normal activity levels; (3) severe-significant dysfunction, such that the patient is unable to perform daily activities; And (4) very severe-the patient's life is at risk from the event. The relationship between adverse events versus administration of TACI-Fc5 can be assessed using the following definitions: (1) likely-causal relationship is very likely clinically / biologically, initiation of adverse effects and administration of TACI-Fc5 There is a plausible time sequence in between; (2) possible-the causal relationship is clinically / biologically plausible, and there is a plausible time sequence between the onset of adverse effects and administration of TACI-Fc5; (3) unlikely-likely not causal, most likely another recorded cause of adverse effects; And (4) irrelevant-a causal relationship can be explicitly excluded, with another reported cause of adverse effects most likely. Medical diseases present in the baseline that do not worsen in severity or frequency during administration of TACI-Fc5 are not considered adverse events. Deterioration of RA is obtained as part of a disease assessment and is not considered a adverse event unless it is possibly or highly related to the administration of TACI-Fc5.

Table 7: Post-Evaluation Evaluation Schedules for Cohorts 1, 3, and 5

D2 D3-5 D8 D15 D29 D43 D57 D71 D85-92 ECG X X X X X VS X X X X X X X X Physical examination X Physical examination ^A X X X X X X ADA ^B X X X X VIS ^C X Routine laboratory tests X X X X X X X ESR X X X X Urine HP / LP X X X X Anti-TACI-Fc5 antibody X PK / PD X X X X X X X X X Biomarker X X X X X X X B cell count (FACS) X X X X X X X Spit puncture X Synovial Biopsy X A: Complaint directed physical examination B: Assessment of disease activity measured by ADA = serum CRP, ESR and urine HP / LP ratio C: VIS = vaccine immunization status

Table 8: Evaluation Schedule after Dosing of Cohorts 2 and 4

D30 D33 D36 D43 D57 D71 D85 D99 D113-119 ECG X X X VS X X X X X Physical examination X Physical examination ^A X X X X ADA ^B X X VIS ^C X Routine laboratory tests X X X X X ESR X X X Urine HP / LP X X X Anti-TACI-Fc5 antibody X PK / PD X X X X X X X X X Biomarker X X X X X X X X X B cell count (FACS) X X X X X X Synovial Biopsy X A: Appeal-induced physical examination B: ADA = assessment of disease activity as measured by serum CRP, ESR and urine HP / LP ratio C: VIS = vaccine immunization status

Table 9: Post-Dose Evaluation Schedules for Cohort 6

D15 D29 D43 D57 D71 D85 D86 D99 D127 D165-173 ECG X X X VS X X X X X X X X X X Physical examination X Physical examination ^A X X X X X X X X X ADA ^B X X X X X X VIS ^C X Routine laboratory tests X X X X X X ESR X X X X Urine HP / LP X X X X Anti-TACI-Fc5 antibody X X PK / PD X X X X X X X X Biomarker X X X X X X X B cell count (FACS) X X X X X X Synovial Biopsy X Spit puncture X A: Appeal-induced physical examination B: ADA = assessment of disease activity as measured by serum CRP, ESR and urine HP / LP ratio C: VIS = vaccine immunization status

The dose, dosing schedule and route of administration of TACI-Fc5 are shown in Table 10. In summary, 72 male and female RA patients aged 18 to 70 years are included. Patients had a definite diagnosis of active moderate to severe RA for more than 6 months, failed for up to 5 DMARDs, and did not use DMARDs other than methotrexate for more than 4 weeks prior to day 1. In addition, patients were RF positive and were not pregnant during the study and up to 3 months after the last administration of TACI-Fc5. These patients are divided into six cohorts. Three of the cohorts consist of eight patients, two of the cohorts consist of 12 patients, and one cohort consists of 24 patients.

Table 10: Dose and Administration of TACI-Fc5

Cohort Drug therapy Route of administration Cohort 1 (70 mg single dose, 8 patients) TACI-Fc5 70 mg or the corresponding placebo on Day 1 1 0.5 ml subcutaneous injection in the morning Cohort 2 (3 × 70 mg repeated doses per month, 12 patients) TACI-Fc5 70 mg or the corresponding placebo on Days 1, 15 and 29 1 0.5 ml subcutaneous injection in the morning Cohort 3 (210 mg single dose, 8 patients) 210 mg of TACI-Fc5 or the corresponding placebo on day 1. One 1.5 ml subcutaneous injection in the morning Cohort 4 (3 × 210 mg repeated doses per month, 12 patients) TACI-Fc5 210 mg or the corresponding placebo on Days 1, 15 and 29 One 1.5 ml subcutaneous injection in the morning Cohort 5 (630 mg single dose, 8 patients) 630 mg of TACI-Fc5 or the corresponding placebo on Day 1 Three 1.5 ml subcutaneous injections in the morning Cohort 6 (7 x 420 mg repeated doses over 3 months, 24 patients TACI-Fc5 420 mg or the corresponding placebo on days 1, 15, 29, 43, 57, 71 and 85 Two 1.5 ml subcutaneous injections in the morning

Example 4

TACI-Fc5 injection procedure.

If a subcutaneous route of administration is chosen to deliver TACI-Fc5, the molecule is injected subcutaneously into the abdominal wall, with the site circulating according to the diagram (Figure 2). Be careful not to inject into the blood vessel. It is extremely important to circulate the area to keep the skin healthy. Repeat injections at the same point can cause scarring and hardening of adipose tissue. The following areas are used for the injection and must be circulated as follows (FIG. 2): first scan into the upper left outer area (FIG. 2, position 1); Secondary scan into the lower right outer region (FIG. 2, position 2), tertiary scan into the lower left outer region (FIG. 2, position 3); Quaternary scan into the upper right outer region (FIG. 2, position 4); And 5 th scan into the median lower region (FIG. 2, position 5). The above mentioned circulation scheme is repeated for further injections.

For patients in need of one or more injections per dose, the injection starts at 12 o'clock on the indicated area of the area (sites 1 to 5 according to FIG. 2) for such injections, and then the number of injections required per dose Are sequentially rotated clockwise to the 2, 4, 6, 8 and / or 10 o'clock position as needed. The injections need to be spaced at least 2.5 cm (1 inch) from each other and are injected as close as possible in time. Injection of more than 1.5 mL into one injection site is not acceptable. If the patient has difficulties with injection into the abdomen, the alternative sites that can be injected are the anterior thigh and the upper arm.

Common symptoms of injection site reactions include itching, tenderness, warmth and / or redness at the injection site.

Example 5

Dose escalation protocol for administering TACI-Ig molecules

This dose escalation protocol for evaluation of TACI-Fc5 in RA enables the administration of 6 patients in the first cohort. After the first dose of 70 mg is administered to the first cohort, observe for 4 weeks. The Safety Review Board (SRB) reviews the safety data collected for 4 weeks after dosing, and at the recommendation of the SRB, the dose is increased to 210 mg over a month (second cohort) or as a single dose. (Third cohort). The second cohort is administered to 9 patients and the third cohort is administered to 6 patients. SRBs then reviewed the safety data collected from the third cohort six weeks after dosing, and at the recommendation of SRB incremented the dose to 630 mg over a month (fourth cohort) or as a single dose (5 First cohort). The fourth cohort is administered to 9 patients and the fifth cohort is administered to 6 patients. The SRB then reviews the safety data collected from the fifth cohort six weeks after administration and administers 2940 mg over a three month period to cohort 6 at the recommendation of the SRB. In the sixth cohort, 18 patients are administered.

The selected dose is reasonably based on repeated SC toxicology studies in cynomolgus monkeys and mice that showed that TACI-Fc5 was well tolerated at 5, 20 and 80 mg / k. Apart from the effect on immune cells, which is the expected result of TACI-Fc5 pharmacokinetic activity, the only result is a significant decrease in plasma total protein and a slight subcutaneous response associated with the highest SC dose of 80 mg / kg in monkeys, and 80 The dose of mg / kg was transient hyperactivity in the Irwin test of mice. Thus, 20 mg / kg is the highest dose not associated with adverse outcomes in both species.

In addition, doses of 2.1, 70, 210 and 630 mg were administered to healthy volunteers as a single subcutaneous dose. No safety or tolerability issues were raised at these doses. Based on preclinical data and preliminary results in healthy volunteers, 70 mg was chosen as the appropriate starting dose for patients with RA.

Dosages and dosing regimens foreseen in RA patients in this study range from 1 mg / kg to 9 mg / kg twice a month (normalized for 70 kg humans), which is the range of doses studied in monkeys. Corresponds well with Repeated dosing in monkeys showed a clear dose-response relationship.

Example 6

Assessment of disease progression.

Disease progression scores include Disease Activity Score, Physician's Global Assessment of Disease Activity, Patient's Global Assessment of Disease Activity, Morning Stiffness, and Measured by patient pain, disease activity and physical function assessment.

Disease Activity Score (DAS) is a combined index developed to measure disease activity in patients with RA. This, together with the EULAR response criteria, has been comprehensively confirmed for its use in clinical trials. The first DAS included an overall health assessment of the Ritchie joint index, 44 swelling joint counts, ESR and VAS. After identification of 28 unranked joint counts for tenderness and swelling, DAS28 was developed comprising 28 joint counts. The results of DAS and DAS28 are not directly compatible.

Using DAS, several thresholds have been developed for high disease activity, low disease activity or even remission. In addition, response criteria have been developed based on DAS, so that the clinical response can be assessed when the patient's DAS is measured at two time points.

Disease variables needed to calculate DAS28 are: swollen joint count (28 joints); Tender joint count (28 joints); ESR (mm after 1 hour); Patient's overall health (GH) or overall disease activity (visual analogue scale (VAS) of 100 mm)-referred to in this study as the assessment of the patient's overall disease activity. The 28 tender and swelling joint scores included the following joints: shoulder, elbow, wrist, metacarpophalangeal joint, proximal interphalangeal joint and knee.

The formula used to calculate the DAS is: DAS28 = 0.56 * sqrt (tension 28) + 0.28 * sqrt (single 28) + 0.70 * ln (ESR) + 0.014 * GH. DAS provides numbers on a scale of 0 to 10, which indicates the current activity of rheumatoid arthritis. DAS28 above 5.1 means high disease activity and DAS28 below 3.2 shows low disease activity. Mitigation is achieved by DAS28 lower than 2.6. When comparing DAS28 from one patient at two different time points, it is possible to define an improvement or response.

The patient's pain assessment and the patient's overall disease activity assessment are performed using the horizontal visual analogy scale (VAS).

The physician's overall disease activity assessment measures the patient's current disease activity using a five-point scale (Likert scale), where 0 = asymptomatic, 1 = mild, 2 = moderate, and 3 = severe And 4 = super severe.

The evaluation of the patient's physical function may be directed to the patient's ability to perform routine activities such as dressing and grooming, eating and walking over the previous week and to assist or implement the patient in performing any of the above-mentioned activities. Measures are based on the patient's response to the Health Assessment Questionnaire (HAQ), which asks the patient as to whether or not he or she needed it.

The presence of morning stiffness is assessed by interviewing the patient and, if present, the duration is recorded.

                         SEQUENCE LISTING <110> Herve, Broly        Busby, Sharon        Gross, jane        Visich, Jennifer        Nestorov, Ivan <120> METHODS FOR TREATING AUTOIMMUNE DISEASES USING A TACI-IG FUSION        MOLECULE <130> 05558.0041.00PC00 <150> US 60 / 747,270 <151> 2006-05-15 <160> 2 <170> PatentIn version 3.4 <210> 1 <211> 154 <212> PRT <213> TACI <400> 1 Met Ser Gly Leu Gly Arg Ser Arg Arg Gly Gly Arg Ser Arg Val Asp 1 5 10 15 Gln Glu Glu Arg Phe Pro Gln Gly Leu Trp Thr Gly Val Ala Met Arg             20 25 30 Ser Cys Pro Glu Glu Gln Tyr Trp Asp Pro Leu Leu Gly Thr Cys Met         35 40 45 Ser Cys Lys Thr Ile Cys Asn His Gln Ser Gln Arg Thr Cys Ala Ala     50 55 60 Phe Cys Arg Ser Leu Ser Cys Arg Lys Glu Gln Gly Lys Phe Tyr Asp 65 70 75 80 His Leu Leu Arg Asp Cys Ile Ser Cys Ala Ser Ile Cys Gly Gln His                 85 90 95 Pro Lys Gln Cys Ala Tyr Phe Cys Glu Asn Lys Leu Arg Ser Pro Val             100 105 110 Asn Leu Pro Pro Glu Leu Arg Arg Gln Arg Ser Gly Glu Val Glu Asn         115 120 125 Asn Ser Asp Asn Ser Gly Arg Tyr Gln Gly Leu Glu His Arg Gly Ser     130 135 140 Glu Ala Ser Pro Ala Leu Pro Gly Leu Lys 145 150 <210> 2 <211> 348 <212> PRT <213> Human Immunoglobulin-Constant Domain Fc5 <400> 2 Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly 1 5 10 15 Ala Val Phe Val Ser Leu Ser Gln Glu Ile His Ala Glu Leu Arg Arg             20 25 30 Phe Arg Arg Ala Met Arg Ser Cys Pro Glu Glu Gln Tyr Trp Asp Pro         35 40 45 Leu Leu Gly Thr Cys Met Ser Cys Lys Thr Ile Cys Asn His Gln Ser     50 55 60 Gln Arg Thr Cys Ala Ala Phe Cys Arg Ser Leu Ser Cys Arg Lys Glu 65 70 75 80 Gln Gly Lys Phe Tyr Asp His Leu Leu Arg Asp Cys Ile Ser Cys Ala                 85 90 95 Ser Ile Cys Gly Gln His Pro Lys Gln Cys Ala Tyr Phe Cys Glu Asn             100 105 110 Lys Leu Arg Ser Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro         115 120 125 Pro Cys Pro Ala Pro Glu Ala Glu Gly Ala Pro Ser Val Phe Leu Phe     130 135 140 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 145 150 155 160 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe                 165 170 175 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro             180 185 190 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr         195 200 205 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val     210 215 220 Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala 225 230 235 240 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg                 245 250 255 Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly             260 265 270 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro         275 280 285 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser     290 295 300 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 305 310 315 320 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His                 325 330 335 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys             340 345  

Claims (14)

A fragment thereof that binds to the TACI extracellular domain or BlyS (i); And A method of treating rheumatoid arthritis in a patient comprising administering to the patient an amount of the composition comprising a fusion molecule comprising human immunoglobulin-constant domain (ii) in an amount effective to treat rheumatoid arthritis. The method of claim 1, wherein said TACI extracellular domain has the sequence set forth in SEQ ID NO: 1. The method of claim 1, wherein said TACI extracellular domain is at least 50% identical to SEQ ID NO: 1. The method of claim 1, wherein said human immunoglobulin-constant domain has the sequence set forth in SEQ ID NO: 2. The method of claim 1, wherein the composition is administered in an amount of about 0.01 mg to about 10 mg per kg body weight of the patient. 6. The method of claim 5, wherein said composition is administered seven times in said amount over a 12 week period. 6. The method of claim 5, wherein after said composition is administered seven times in said amount for a 12 week period, said composition is further administered in said amount. 6. The method of claim 5, wherein said composition is administered three times in said amount for a four week period. 6. The method of claim 5, wherein after said composition is administered three times in said amount for a four week period, said composition is further administered in said amount. The method of claim 5, wherein the composition is administered biweekly in the amount for 2 to 30 weeks. The method of claim 1, wherein the method further comprises co-administering a second agent to the patient. 12. The method of claim 11, wherein the second agent is hydroxychloroquine, sulfasalazine, methotrexate, leflunomide, rituximab, infliximab, azathioprine, D-penicillamine, Gold (oral or Root), minocycline, cyclosporin, corticosteroids, non-steroidal anti-inflammatory drugs (NSAIDs), cytokines, anti-cytokines and interferons. The method of claim 1, wherein the composition is administered subcutaneously, orally or intravenously. The method of claim 1, wherein the patient is a human.

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